Silymarin induces inhibition of growth and apoptosis through modulation of the MAPK signaling pathway in AGS human gastric cancer cells

Kim,Sung‑Hyun; Choo,Gang‑Sik; Yoo,Eun‑Seon; Woo,Joong‑Seok; Han,So‑Hee; Lee,Jae‑Han; Jung,Ji‑Youn

doi:10.3892/or.2019.7295

Silymarin induces inhibition of growth and apoptosis through modulation of the MAPK signaling pathway in AGS human gastric cancer cells

Authors:
- Sung‑Hyun Kim
- Gang‑Sik Choo
- Eun‑Seon Yoo
- Joong‑Seok Woo
- So‑Hee Han
- Jae‑Han Lee
- Ji‑Youn Jung
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Affiliations: Department of Companion and Laboratory Animal Science, Kongju National University, Yesan 340‑702, Republic of Korea
Published online on: August 28, 2019 https://doi.org/10.3892/or.2019.7295
Pages: 1904-1914
Copyright: © Kim et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Apoptosis is regarded as a therapeutic target because it is typically disturbed in human cancer. Silymarin from milk thistle (Silybum marianum) has been reported to exhibit anticancer properties via regulation of apoptosis as well as anti‑inflammatory, antioxidant and hepatoprotective effects. In the present study, the effects of silymarin on the inhibition of proliferation and apoptosis were examined in human gastric cancer cells. The viability of AGS human gastric cancer cells was assessed by MTT assay. The migration of AGS cells was investigated by wound healing assay. Silymarin was revealed to significantly decrease viability and migration of AGS cells in a concentration‑dependent manner. In addition, the number of apoptotic bodies and the rate of apoptosis were increased in a dose‑dependent manner as determined by DAPI staining and Annexin V/propidium iodide double staining. The changes in the expression of silymarin‑induced apoptosis proteins were investigated in human gastric cancer cells by western blotting analysis. Silymarin increased the expression of Bax, phosphorylated (p)‑JNK and p‑p38, and cleaved poly‑ADP ribose polymerase, and decreased the levels of Bcl‑2 and p‑ERK1/2 in a concentration‑dependent manner. The in vivo tumor growth inhibitory effect of silymarin was investigated. Silymarin (100 mg/kg) significantly decreased the AGS tumor volume and increased apoptosis, as assessed by the TUNEL assay, confirming its tumor‑inhibitory effect. Immunohistochemical staining revealed elevated expression of p‑JNK and p‑p38 as well as reduced expression of p‑ERK1/2 associated with silymarin‑treatment. Silymarin was revealed to reduce tumor growth through inhibition of p‑ERK and activation of p‑p38 and p‑JNK in human gastric cancer cells. These results indicated that silymarin has potential for development as a cancer therapeutic due to its growth inhibitory effects and induction of apoptosis in human gastric cancer cells.

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1	Jung KW, Won YJ, Kong HJ and Lee ES: Prediction of cancer incidence and mortality in Korea, 2019. Cancer Res Treat. 51:431–437. 2019. View Article : Google Scholar : PubMed/NCBI
2	Won YJ, Sung J, Jung KW, Kong HJ, Park S, Shin HR, Park EC, Ahn YO, Hwang IK, Lee DH, et al: Nationwide cancer incidence in Korea, 2003–2005. Cancer Res Treat. 41:122–131. 2009. View Article : Google Scholar : PubMed/NCBI
3	Jung KW, Won YJ, Kong HJ, Oh CM, Lee DH and Lee JS: Cancer statistics in Korea: Incidence, mortality, survival, and prevalence in 2011. Cancer Res Treat. 46:109–123. 2014. View Article : Google Scholar : PubMed/NCBI
4	Jemal A, Center MM, DeSantis C and Ward EM: Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomarkers Prev. 19:1893–1907. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
6	Forman D and Burley VJ: Gastric cancer: Global pattern of the disease and an overview of environmental risk factors. Best Pract Res Clin Gastroenterol. 20:633–649. 2006. View Article : Google Scholar : PubMed/NCBI
7	Lee MS, Cha EY, Thuong PT, Kim JY, Ahn MS and Sul JY: Down-regulation of human epidermal growth factor receptor 2/neu oncogene by corosolic acid induces cell cycle arrest and apoptosis in NCI-N87 human gastric cancer cells. Biol Pharm Bull. 33:931–937. 2010. View Article : Google Scholar : PubMed/NCBI
8	Li N, Fan LL, Sun GP, Wan XA, Wang ZG, Wu Q and Wang H: Paeonol inhibits tumor growth in gastric cancer in vitro and in vivo. World J Gastroenterol. 16:4483–4490. 2010. View Article : Google Scholar : PubMed/NCBI
9	Zhang L, Hou YH, Wu K, Zhai JS and Lin N: Proteomic analysis reveals molecular biological details in varioliform gastritis without helicobacter pylori infection. World J Gastroenterol. 16:3664–3673. 2010. View Article : Google Scholar : PubMed/NCBI
10	Seeram NP: Berry fruits for cancer prevention: Current status and future prospects. J Agric Food Chem. 56:630–635. 2008. View Article : Google Scholar : PubMed/NCBI
11	An IJ, Kwon JK, Lee JS, Park HS, Kim DC, Choi BJ, Lee KM, Park YJ and Jung JY: Induction of apoptosis in human cancer cells with compositae extracts. J Korean Soc Food Sci Nutr. 41:584–590. 2012. View Article : Google Scholar
12	Kwon MJ and Nam TJ: A polysaccharide of the marine alga capsosiphon fulvescens induces apoptosis in AGS gastric cancer cells via an IGF-IR-mediated PI3K/Akt pathway. Cell Biol Int. 31:768–775. 2007. View Article : Google Scholar : PubMed/NCBI
13	Schuier M, Sies H, Illek B and Fischer H: Cocoa-related flavonoids inhibit CFTR-mediated chloride transport across T84 human colon epithelia. J Nutr. 135:2320–2325. 2005. View Article : Google Scholar : PubMed/NCBI
14	Murakami A, Ashida H and Terao J: Multitargeted cancer prevention by quercetin. Cancer Lett. 269:315–325. 2008. View Article : Google Scholar : PubMed/NCBI
15	Ren W, Qiao Z, Wang H, Zhu L and Zhang L: Flavonoids: Promising anticancer agents. Med Res Rev. 23:519–534. 2003. View Article : Google Scholar : PubMed/NCBI
16	Hackett ES, Twedt DC and Gustafson DL: Milk thistle and its derivative compounds: A review of opportunities for treatment of liver disease. J Vet Intern Med. 27:10–16. 2013. View Article : Google Scholar : PubMed/NCBI
17	Chen CH, Huang TS, Wong CH, Hong CL, Tsai YH, Liang CC, Lu FJ and Chang WH: Synergistic anti-cancer effect of baicalein and silymarin on human hepatoma HepG2 Cells. Food Chem Toxicol. 47:638–644. 2009. View Article : Google Scholar : PubMed/NCBI
18	Ramasamy K and Agarwal R: Multitargeted therapy of cancer by silymarin. Cancer Lett. 269:352–362. 2008. View Article : Google Scholar : PubMed/NCBI
19	Surai PF: Silymarin as a natural antioxidant: An overview of the current evidence and perspectives. Antioxidants (Basel). 4:204–247. 2015. View Article : Google Scholar : PubMed/NCBI
20	Katiyar SK: Silymarin and skin cancer prevention: Anti-inflammatory, antioxidant and immunomodulatory effects (Review). Int J Oncol. 26:169–176. 2005.PubMed/NCBI
21	Féher J and Lengyel G: Silymarin in the prevention and treatment of liver diseases and primary liver cancer. Curr Pharm Biotechnol. 13:210–217. 2012. View Article : Google Scholar : PubMed/NCBI
22	Eo HJ, Park GH and Jeong JB: Inhibition of Wnt signaling by silymarin in human colorectal cancer cells. Biomol Ther (Seoul). 24:380–386. 2016. View Article : Google Scholar : PubMed/NCBI
23	Hajighasemlou S, Farajollahi M, Alebouyeh M, Rastegar H, Manazi MT, Mirmoghtadaei M, Moayedi B, Ahmadzadeh M, Kazemi M, Parvizpour F and Gharibzadeh S: Study of the effect of silymarin on viability of breast cancer cell lines. Adv Breast Cancer Res. 3:100–105. 2014. View Article : Google Scholar
24	Wu T, Liu W, Guo W and Zhu X: Silymarin suppressed lung cancer growth in mice via inhibiting myeloid-derived suppressor cells. Biomed Pharmacother. 81:460–467. 2016. View Article : Google Scholar : PubMed/NCBI
25	Deep G, Singh RP, Agarwal C, Kroll DJ and Agarwal R: Silymarin and silibinin cause G1 and G2-M cell cycle arrest via distinct circuitries in human prostate cancer PC3 cells: A comparison of flavanone silibinin with flavanolignan mixture silymarin. Oncogene. 25:1053–1069. 2006. View Article : Google Scholar : PubMed/NCBI
26	Hsu HF, Houng JY, Kuo CF, Tsao N and Wu YC: Glossogin, a novel phenylpropanoid from glossogyne tenuifolia, induced apoptosis in A549 lung cancer cells. Food Chem Toxicol. 46:3785–3791. 2008. View Article : Google Scholar : PubMed/NCBI
27	Kaufmann T, Strasser A and Jost PJ: Fas death receptor signalling: Roles of Bid and XIAP. Cell Death Differ. 19:42–50. 2012. View Article : Google Scholar : PubMed/NCBI
28	Tummers B and Green DR: Caspase-8: Regulating life and death. Immunol Rev. 277:76–89. 2017. View Article : Google Scholar : PubMed/NCBI
29	Kim D and Chung J: Akt: Versatile mediator of cell survival and beyond. J Biochem Mol Biol. 35:106–115. 2002.PubMed/NCBI
30	Burz C, Berindan-Neagoe I, Balacescu O and Irimie A: Apoptosis in cancer: Key molecular signaling pathways and therapy targets. Acta Oncol. 48:811–821. 2009. View Article : Google Scholar : PubMed/NCBI
31	Soldani C and Scovassi AI: Poly (ADP-ribose) polymerase-1 cleavage during apoptosis: An update. Apoptosis. 7:321–328. 2002. View Article : Google Scholar : PubMed/NCBI
32	Kim EK and Choi EJ: Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta. 1802:396–405. 2010. View Article : Google Scholar : PubMed/NCBI
33	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
34	Ramakrishnan G, Lo Muzio L, Elinos-Báez CM, Jagan S, Augustine TA, Kamaraj S, Anandakumar P and Devaki T: Silymarin inhibited proliferation and induced apoptosis in hepatic cancer cells. Cell Prolif. 42:229–240. 2009. View Article : Google Scholar : PubMed/NCBI
35	Zhong X, Zhu Y, Lu Q, Zhang J, Ge Z and Zheng S: Silymarin causes caspases activation and apoptosis in K562 leukemia cells through inactivation of Akt pathway. Toxicology. 227:211–216. 2006. View Article : Google Scholar : PubMed/NCBI
36	Fan L, Ma Y, Liu Y, Zheng D and Huang G: Silymarin induces cell cycle arrest and apoptosis in ovarian cancer cells. Eur J Pharmacol. 743:79–88. 2014. View Article : Google Scholar : PubMed/NCBI
37	Vaid M, Prasad R, Sun Q and Katiyar SK: Silymarin targets β-catenin signaling in blocking migration/invasion of human melanoma cells. PLoS One. 6:e230002011. View Article : Google Scholar : PubMed/NCBI
38	Danial NN and Korsmeyer SJ: Cell death: Critical control points. Cell. 116:205–219. 2004. View Article : Google Scholar : PubMed/NCBI
39	Halicka HD, Bedner E and Darzynkiewicz Z: Segregation of RNA and separate packaging of DNA and RNA in apoptotic bodies during apoptosis. Exp Cell Res. 260:248–256. 2000. View Article : Google Scholar : PubMed/NCBI
40	Katiyar SK, Roy AM and Baliga MS: Silymarin induces apoptosis primarily through a p53-dependent pathway involving Bcl-2/Bax, cytochrome c release, and caspase activation. Mol Cancer Ther. 4:207–216. 2005.PubMed/NCBI
41	Vaid M, Singh T, Prasad R and Katiyar SK: Silymarin inhibits melanoma cell growth both in vitro and in vivo by targeting cell cycle regulators, angiogenic biomarkers and induction of apoptosis. Mol Carcinog. 54:1328–1339. 2015. View Article : Google Scholar : PubMed/NCBI
42	Yan C, Yahua WU, Wang Z, Yin C and Yin M: Inhibitory effect of silymarin on human osteosarcoma Saos-2 cells and its mechanism. Chin Pharmacol Bull. 32:966–969. 2016.(In Chinese).
43	Huang Q, Wu LJ, Tashiro S, Onodera S, Li LH and Ikejima T: Silymarin augments human cervical cancer HeLa cell apoptosis via P38/JNK MAPK pathways in serum-free medium. J Asian Nat Prod Res. 7:701–709. 2005. View Article : Google Scholar : PubMed/NCBI
44	Won DH, Kim LH, Jang B, Yang IH, Kwon HJ, Jin B, Oh SH, Kang JH, Hong SD, Shin JA and Cho SD: In vitro and in vivo anti-cancer activity of silymarin on oral cancer. Tumor Biol. 40:10104283187761702018. View Article : Google Scholar
45	Singh RP, Tyagi AK, Zhao J and Agarwal R: Silymarin inhibits growth and causes regression of established skin tumors in SENCAR mice via modulation of mitogen-activated protein kinases and induction of apoptosis. Carcinogenesis. 23:499–510. 2002. View Article : Google Scholar : PubMed/NCBI